| Objective ① Screening the anti-allergic active constituent from Sophora flavescens Ait.and analyzing its chemical composition.② Study the anti-allergic effects and toxicity of OMT by a variety of different anti-allergic models in vivo and vitro,and discuss the action mechanism.③ The preparation of Anti-allergic nose drops of OMT.④Establish it’s quality standard(draft)and investigate it’s preliminary stability.Methods ① Separating and analyzing of anti-allergic active constituent:Using solvent extraction and silica gel column chromatography combined with the anti-allergic experimental to screen the anti-allergic active constituent and analyzing its chemical composition by TLC and HPLC.② Anti-allergic effects research:evaluating the anti-allergic effects on guinea pig ileum experiments,passive cutaneous anaphylaxis reaction of ICR mice,allergic rhinitis model of mice(by oral and nasal drops administration)and rats.And discuss the anti-allergic mechanism by determining the concentration of histamine,NO,IgE and EOS in the serum.Acute toxicity studies on ICR mice,determining LD50 to evaluat its safety.③ The preparation of anti-allergic nasal drops:Investigating the pH and viscosity of nasal drops and best technique condition was determined.According to the prescription,prepare three batch samples for the validation experiments.④Anti-allergic nasal drops’s quality standards:Formulate quality standard(draft)of the appearance,TLC,content determination,pH,clarity.⑤ The preliminary stability test:Examining the preliminary stability in the 0 to 3months(once a month)by investigating the appearance,TLC,content determination,pH,clarity at room temperature.Results ① The active constituent of anti-allergic from Sophora flavescens Ait.was OMT and its content and purity is 0.559%and 95.58%。②OMT can inhibit the contraction of isolated guinea pig ileum caused by histamine(p<0.05)obviously.And the strongest inhibition rate is 52.34%.③ High and low dose group of OMT reduced significantly the permeability of blood vessel and decrease the leakage of Evans blue in the Passive Cutaneous Anaphylaxis reaction(p<0.05).The inhibition rate is 57.14%and 50.00%respectively.④Method of administration by oral:High and low dose group of OMT improved significantly mice allergic symptoms of allergic rhinitis induced by OVA(p<0.05).The ELISA experiment showed high dose group can reduce significantly the content of histamine and NO in the serum,but low dose group not obviously declined.High dose group’s anti-allergic effects are consistent with Loratadine group’s.⑤ Method of administration by nasal drops:High middle and low dose group of OMT improved significantly mice allergic symptoms of allergic rhinitis induced by OVA(p<0.05).The ELISA experiment showed high middle and low dose group can reduce significantly the content of histamine and NO in the serum.⑥High and low dose group of OMT improved significantly rats allergic symptoms of allergic rhinitis induced by OVA(p<0.05).The number of EOS was decreased significantly compared with the model group.The ELISA experiment showed high dose and low dose group can reduce significantly the content of histamine and IgE in the serum.Rat nasal mucosal tissue using the test results showed that different dosage group compared with model group,lymphocytes and plasma cells were decreased,table ciliated columnar epithelium form basic returned to normal.⑦ According to the result of acute toxicity studies on ICR mice,LD50 of OMT is 668.565mg·kg-1.⑧Preparation of prescription of anti-allergic nasal drops:OMT power 50g,NaCl 9g,CMC-Na 3g,thiomersalate 0.1g,and use 1%citrate to adjust the pH,refrigerate for 48 hours and split charging.⑨Anti-allergic nasal drops are polymer solution which is slightly sticky,transparent and stable.According to thin layer identification,the tablets have the same points with OMT standard substance.⑩The test results of appearance,TLC,content determination,pH,clarity,disintegration time of 1,2,3 months anti-allergic nasal drops,compared with 0 month,these indicators have no obvious changes.Conclusion ① The active constituent of anti-allergic from Sophora flavescens Ait.was OMT.②OMT has obvious inhibitory effects on guinea pig ileum experiments,passive cutaneous anaphylaxis reaction and allergic rhinitis model of mice and rats.③The anti-allergic effects may be related to decline of the content of histamine NO and IgE in the allergic rhinitis in mice and rats induced by OVA.The rat nasal mucosa biopsy can prompt bitter and alkali oxide by reducing nasal mucosal epithelium infiltrating lymphocytes and plasma cells,reduce allergic media release,so as to achieve the effect of treatment of allergic rhinitis.④ The preparation process of anti-allergic nasal drops is reasonable and feasible.Its quality can be well controlled and it has good stability from appearance to inner quality. |
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